The effects of bisphosphonates on MMP-9 activity and migration of different PC3 cells. A. The effect of BPs, such as alendronate (AL, lanes 3, 6, and 9) and pamidronate (PA, lanes 4, 7, and 10) on MMP-9 activity is shown by gelatin zymography. MMP-9 activity was determined in the conditioned media of different PC3 cells indicated in the figure. Untreated cells were used as controls (lanes 2, 5, and 8). The activity of a recombinant MMP-9 protein was used as an identification control. Arrows indicate pro- and active forms of MMP-9. The results represent one of three separate experiments performed. B. The effect of BP on the total cellular levels of MMP-9 was determined by immunoblotting with an antibody to MMP-9. No significant changes in the total cellular levels of MMP-9 were observed in response to BP treatment. C-H. The effects of pamidronate (PA) and GM6001 (GM) on the migration of PC3 cell lines are shown by a wound closure assay. Statistical analysis is provided at the bottom of each panel as unhealed wound distance (in μm) at 48 h with (+) and without (-) pamidronate (panels E and F) or GM6001 (panels G and H). The data represent the mean ± SEM of triplicate determinations of three different experiments. I. The effects of pamidronate on the proliferation of PC3 cells. Dotted line indicates the initial cell number at the time of plating (2.5 × 104 cells). PC3 cells were treated with 10–100 μM PA for 48 h. A dose dependent decrease in the proliferation was observed in response to PA treatment. Data represent a total of three independent experiments and are expressed as mean ± SEM.